Mengaud J, Vicente M F, Chenevert J, Pereira J M, Geoffroy C, Gicquel-Sanzey B, Baquero F, Perez-Diaz J C, Cossart P
Unité de Génie Microbiologique, Institut Pasteur, Paris, France.
Infect Immun. 1988 Apr;56(4):766-72. doi: 10.1128/iai.56.4.766-772.1988.
To evaluate the role of hemolysin production in the virulence of Listeria monocytogenes, we have undertaken the analysis of the chromosomal region containing hlyA, the gene coding for listeriolysin O. A recombinant cosmid, conferring a hemolytic phenotype to Escherichia coli, was shown to express listeriolysin O, by immunoblotting with a specific antiserum against listeriolysin O. The presence of hlyA on the cosmid was demonstrated by DNA hybridization with a probe previously shown to contain part of hlyA. The complete nucleotide sequence of hlyA has been determined. The deduced protein sequence reveals the presence of a putative 25-amino-acid signal sequence: the secreted form of listeriolysin O would have 504 amino acids, in agreement with the molecular weight of purified listeriolysin O (58,000). The protein sequence is highly homologous to those of streptolysin O and pneumolysin. A peptide of 11 amino acids conserved in the three proteins contains the unique cysteine known to be essential for lytic activity. By DNA-DNA hybridization, the listeriolysin O gene was detected in all L. monocytogenes strains tested, even in the nonhemolytic type strain. The gene was absent in other species of the genus Listeria.
为了评估溶血素产生在单核细胞增生李斯特菌毒力中的作用，我们对包含hlyA（编码李斯特菌溶血素O的基因）的染色体区域进行了分析。通过用针对李斯特菌溶血素O的特异性抗血清进行免疫印迹，显示赋予大肠杆菌溶血表型的重组黏粒表达了李斯特菌溶血素O。通过与先前显示包含hlyA部分的探针进行DNA杂交，证实了黏粒上存在hlyA。已确定hlyA的完整核苷酸序列。推导的蛋白质序列显示存在一个推定的25个氨基酸的信号序列：李斯特菌溶血素O的分泌形式将有504个氨基酸，这与纯化的李斯特菌溶血素O的分子量（58,000）一致。该蛋白质序列与链球菌溶血素O和肺炎溶血素的序列高度同源。在这三种蛋白质中保守的一个11个氨基酸的肽段包含已知对裂解活性至关重要的独特半胱氨酸。通过DNA-DNA杂交，在所有测试的单核细胞增生李斯特菌菌株中都检测到了李斯特菌溶血素O基因，甚至在非溶血型菌株中也有。在李斯特菌属的其他物种中未发现该基因。
